We are requesting fund to purchase an Amis Image Stream, a unique instrument that combines flow cytometry with optical imaging to create a rea- sonbly detailed representation of each cell that passes through the chamber. This image is assembled from multiple image stripes captured as the cell passes across CCD- recording chips. There are six detection channels;two for bright and dark field, respect- timely and four for fluorescence. The fluorescence is detected from excitation via 3 well- separated lasers (405, 488, and 658 in the requested instrument) and the image Chan- nels are then analyzed separately. This instrument will be installed in the Yale FACS Core, in publicly accessible dedicated space. The FACS Core currently services over 200 PIs and 1100 individual users. Fifteen PIs were identified as major users and seven as minor users of the new instrument and their combined planned use alone will more than account for the available hours on the new machine. These PIs have over $17M in NIH-funded direct costs from at least 6 different institutes. While naturally a major re- search focus is in the broad area of immunology and allied immune diseases, disciplines such as stem cell biology, hematopoiesis, infectious disease, vascular biology are well- represented. While the FACS Core is well-endowed with traditional (though busy) ana- lyzers and sorters, the Image Stream brings a totally new dimension. Our extensive pre- liminary and demo work proved to us that this instrument will truly allow our investigators to break new ground in their research. The demo phase of this instrument indeed crew- ated great excitement among the PIs and hands-on users. The new machine will be well cared for by the staff of the FACS Core, which has an extensive infrastructure and culture in place to train users, oversee the machine and ensure that it is self-supporting. Two different technicians will be responsible for the conceptual and day to day aspects of experiments on the Image Stream. We will start and certify the machine each morning and troubleshoot any problems, as we will do at any time during working hours. As such, it will have a remarkable impact on the productivity of an outstanding and well-funded group of investigators. PUBLIC HEALTH RELEVANCE: This instrument will enable the research of 21 NIH-funded principal investigators to enter a new area that combines image-analysis and flow cytometry using the unique Amnis ImageStream analyzer. There is no such existing machine at Yale or even close by. A wide variety of immunology, cell biology, cancer and stem cell research can take advantage of this new approach;our users have $17M per year of NIH funded research that will benefit.